Methods for alleviating, inhibiting or reversing muscle disuse atrophy in mammals

ABSTRACT

Methods for alleviating one or more symptoms and/or inhibiting muscle disuse atrophy in mammals via administration of compositions of egg powder protein are provided.

This patent application claims the benefit of priority from U.S.Provisional Application Ser. No. 62/790,612 filed Jan. 10, 2019, U.S.Provisional Application Ser. No. 62/756,279 filed Nov. 6, 2018, and U.S.Provisional Application Ser. No. 62/724,263, filed Aug. 29, 2018, theteachings of each of which are herein incorporated by reference in theirentireties.

BACKGROUND

Muscle loss due to muscle disuse atrophy is a common problem thatmammals experience following many common orthopedic surgical procedures.

Skeletal muscle atrophy is characterized by a decrease in proteincontent, fiber diameter, force production and resistance to fatigue. Inmuscle disuse atrophy, a decreased rate of protein synthesis along withan increase in protein degradation, also referred to as proteolysis,have been shown to be important contributing factors (Jackman, R W. andKandarian, S. C. American Journal of Physiology-Cell Physiology 2004287(4):C834-C843). In this 2004 review article, Jackman and Kandariandiscuss the interaction of molecular pathways includingubiquitin-proteasome, lysosomal and calpain in muscle proteolysis duringmuscle atrophy (American Journal of Physiology-Cell Physiology 2004287(4):C834-C843). Under specific conditions, TNF-α, glucocorticoids,myostatin and reactive oxygen species can induce muscle atrophy. Inmuscle disuse atrophy, the transcription factor NF-κB plays aparticularly important role. The key physiological stimulus in muscledisuse atrophy is when a muscle no longer bears weight (Goldspink et al.Pflügers Archiv 1986 407(3):333-340). The removal of weight bearinginitiates a cascade of molecular signaling events.

Total hip arthroplasty (THA) is a common orthopedic surgery procedurethat is performed to relieve pain and improve functional performance inhumans with end-stage hip osteoarthritis. THA is typically followed byout-patient physical rehabilitation. However, it has been widelyreported that many patients experience muscle atrophy and reduced musclestrength and functional performance at various time-points followingtheir THA surgical procedure (Holm et al. PloS One 2013 8(4):e62109;Holstege et al. Archives of Physical Medicine and Rehabilitation 201192(2):236-241; Jensen et al. Osteoarthritis and Cartilage 201119(9):1108-1116). According to Freeman et al. (Clinical Biomechanics2013 28(2):171-177), the failure of the central nervous system (CNS) toactivate muscles close to the operated hip joint as a result ofintra-articular swelling may play a role.

A similar form of muscle atrophy referred to as arthrogenic muscleinhibition is commonly observed after total knee arthroplasty (TKA)(Mizneret al. The Journal of Bone and Joint Surgery 2005 87(5):1047;Rice, D. A. and McNair P. J. Seminars in Arthritis and Rheumatism. Vol.40. No. 3. WB Saunders, 2010).

The anterior cruciate ligament (ACL) is a ligament that connects thetibia to the femur. An injury of this ligament typically involves theACL becoming stretched, partially torn or completely torn. In order torepair the ACL, a surgical procedure known as ACL reconstruction surgeryis performed and the surgery involves replacing the torn ACL with agraft prepared from a tendon that is taken from another source such asthe patellar tendon, hamstring tendon or quadriceps tendon. The graftserves as a scaffolding, upon which a new ligament tissue will grow.Researchers have reported a statistically significant loss of musclemass and muscle strength in the quadriceps six months following surgery.

Similar muscle loss is observed in dogs following orthopedic surgery.

For example, Tibial-Plateau-Leveling Osteotomy (TPLO) surgery is aveterinary orthopedic surgical procedure that is commonly performed ondogs in order to stabilize their stifle joint after a rupture of thecranial cruciate ligament (CCL). The CCL is analogous to the anteriorcruciate ligament (ACL) in humans. In dogs that undergo TPLO surgery,the CCL typically ruptures due to long-term degeneration as the fibersthat comprise the ligament undergo degradation over a long period oftime. Although the exact cause of the degeneration of the CCL is notfully understood, genome wide association studies (GWAS) have providedsome insight into the genetic predisposition for CCL degeneration withthe identification of single nucleotide polymorphisms (SNPs) identifiedin the SORCS2 and SEMA5B genes that are thought to play a role (Baird etal. Animal Genetics 2014 45(4):542-549). Certain breeds of dogs such asLabradors, Rottweilers, Boxers, West Highland White Terriers andNewfoundlands are believed to be genetically predisposed to CCL rupture.In addition to genetics, factors such as obesity (Adams et al. Journalof Small Animal Practice 2011 52(7):347-352), individual conformation(Guerrero et al. American Journal of Veterinary Research 200768(12):1332-1337), hormonal imbalance (Light et al. American Journal ofVeterinary Research 2012 73(8):1186-1193) and inflammatory conditions(Lemburg et al. Veterinary Immunology and Immunopathology 200497(3-4):231-240) that impact the joint are believed to exacerbate thedegeneration of the CCL. The CCL traverses from the cranial mid part ofthe tibial intercondylar eminence to the lateral condyle of the femur.Under regular circumstances, the CCL prevents backward movement of thefemur with respect to the tibia. However, due to selective breeding ofdogs, the tibial plateau slope has sloped too far backwards which placesconstant stress on the CCL. This constant stress over time eventuallyleads to a degenerative rupture of the CCL. Rupture of the CCL is verypainful for the dog and typically leads to chronic progressive arthritisin the stifle if not properly treated. During the TPLO surgicalprocedure, the portion of the tibia that adjoins the stifle known as thetibial plateau, is cut by the surgeon and rotated so that its slopechanges to approximately 5 degrees with respect to the horizontal plane.The surgical procedure prevents the femur from sliding down the slope ofthe tibial plateau where the canine exerts its weight on its knee. Afterundergoing TPLO surgery, the majority of dogs regain an active lifestylewithout suffering from many post-operative complications and requiringdrugs for long-term pain management. However, muscle loss due to atrophyof immobilization is a problem that dogs typically encounter after aTPLO surgical procedure (Monk et al. American Journal of VeterinaryResearch 2006 67(3):529-536).

There is a need for compositions and methods of alleviating muscledisuse atrophy in mammals.

SUMMARY

The present invention relates to a method for inhibiting, reversing oralleviating muscle disuse atrophy in a mammal via administration of acomposition comprising egg yolk powder to the mammal.

In one nonlimiting embodiment, the composition administered isFORTETROPIN.

In one nonlimiting embodiment, the composition is administered to themammal following a surgical procedure. In one nonlimiting embodiment,the surgical procedure is an orthopedic surgical procedure.

In one nonlimiting embodiment, the composition is administered to themammal prior to and following a surgical procedure. In one nonlimitingembodiment, the surgical procedure is an orthopedic surgical procedure.

In one nonlimiting embodiment, the composition is administered to themammal following an injury causing immobilization.

In the present invention, administration of the composition after orbefore and after the surgical procedure or after the immobilizing injurycan result in reduced muscle atrophy of disuse, lower reduction inmuscle thickness and/or muscle mass, improved muscle function, increasedmuscle force generation, reduced inflammation, reduced recovery timeand/or lower increases in myostatin in the mammal.

DETAILED DESCRIPTION

The present invention provides methods and compositions for inhibitingand/or reversing and/or alleviating muscle disuse atrophy in a mammal.

Compositions administered in accordance with the present inventioncomprise egg yolk powder or one or more proteins and/or lipids derivedfrom egg yolk which are effective in growth and development of leanmuscle tissue.

In one nonlimiting embodiment, the composition administered isFORTETROPIN. FORTETROPIN is a fertilized egg yolk derived product usedas a dietary and nutritional supplement (MYOS RENS TECHNOLOGY INC.CORPORATION Cedar Knolls, N.J.). A method for production of FORTETROPINis disclosed in U.S. Pat. No. 8,815,320, teachings of which are hereinincorporated by reference in their entirety.

In another nonlimiting embodiment, the composition comprises an avianfollistatin such as described in U.S. Published Patent Application No.2007/0275036, the disclosure of which is incorporated herein byreference in its entirety and/or other proteins and/or lipids found inavian eggs and which are beneficial in growth and development of leanmuscle tissue such as, but not limited to, those described in U.S.Provisional Application Ser. No. 62/755,601, the disclosure of which isincorporated herein by reference in its entirety.

For purposes of the present invention, when the phrases “egg yolkpowder” or “composition comprising egg yolk powder” are used, they aremeant to be inclusive, but are not limited to, egg yolk powder, one ormore proteins and/or lipids derived from egg yolk which are effective ingrowth and development of lean muscle tissue, FORTETROPIN and avianfollistatin.

Disclosed herein are methods for use of compositions comprising egg yolkpowder in the alleviation, reversal and/or inhibition of muscle disuseatrophy in mammals. In these methods, the composition is administered toa mammal in need thereof in an amount effective to alleviate one or moresymptoms of muscle disuse atrophy and/or reverse muscle disuse atrophyand/or inhibit muscle disuse atrophy in the mammal.

By “mammal” or “mammals” it is meant to include, but is not limited to,humans, apes, monkeys, cows, sheep, goats, dogs, cats, mice, rats, andtransgenic species thereof.

Nonlimiting examples of mammals “in need” for purposes of the presentinvention are mammals that have undergone as well mammals that arescheduled to undergo a surgical procedure and mammals that have sufferedan immobilizing injury. In one nonlimiting embodiment, the surgicalprocedure or immobilizing injury is expected to result in muscle disuseatrophy. In one nonlimiting embodiment, the surgical procedure is anorthopedic surgical procedure. Nonlimiting examples of orthopedicsurgical procedures in which the composition comprising egg yolk powdercan be administered after or before and after the procedure includeTPLO, TKA, THA and ACL reconstruction surgery. Nonlimiting examples ofimmobilizing injuries in which the composition comprising egg yolkpowder can be administered after include sports injuries, broken bonesand sprained ankles.

By “alleviation and/or reversal and/or inhibition of muscle disuseatrophy” for purposes of this invention, it is meant to include, but isnot limited to, reduced muscle atrophy of disuse, lower reduction inmuscle thickness and/or muscle mass, improved muscle function, increasedmuscle force generation, reduced inflammation, reduced recovery timeand/or lower increases in myostatin, a protein which can induce muscleatrophy, in the mammal.

In one nonlimiting embodiment, the composition administered isFORTETROPIN. In one nonlimiting embodiment, the FORTETROPIN is producedin accordance with the process described in U.S. Pat. No. 8,815,320.

In one nonlimiting embodiment, FORTETROPIN production is optimized toenhance potency as it relates to muscle disuse atrophy by modifying oneor more egg yolk-related parameters such as, but not limited to,incubation time post-lay, fertility status and breed of chicken.

In one nonlimiting embodiment, the composition comprising egg yolkpowder is administered orally on a daily basis, one, two or three timesa day.

In one nonlimiting embodiment, the composition comprising egg yolkpowder is administered in an amount effective to upregulate mTor pathwayactivity, downregulate ubiquitin proteasome pathwayactivity,downregulate serum myostatin levels and/or reduce ActRIIBexpression, a receptor for myostatin.

In one nonlimiting embodiment, the composition comprising egg yolkpowder is FORTETROPIN administered orally on a daily basis in an amountranging from about 5 to about 25 grams/day, about 6.6 to about 19.8grams/day or about 50 to about 300 mg/kg/day, about 80 to about 250mg/kg/day in humans. In canines, FORTETROPIN is administered orally on adaily basis in an amount ranging from about 200-1000 mg/kg/day, about300 to about 900 mg/kg/day.

In one nonlimiting embodiment, administration of the compositioncomprising egg yolk powder is started about 1 to about 4 weeks beforesurgery.

In one nonlimiting embodiment, administration of the compositioncomprising egg yolk powder occurs after surgery and continues for about2 to 12 weeks, more preferably 6 to 12 weeks.

In one nonlimiting embodiment, administration of the compositioncomprising egg yolk powder is started before surgery and is continuedafter surgery. In one nonlimiting embodiment, administration of thecomposition comprising egg yolk powder is started about 1 to about 4weeks before surgery and continues after surgery for at least 2 to 12weeks.

In one nonlimiting embodiment, the composition comprising egg yolkpowder is administered in combination with one or more additional agentswhich alleviate pain and/or inflammation post surgery or post injuy.Nonlimiting examples of additional agents which can be administered incombination with a composition comprising egg yolk powder include, butare not limited to, curcumin, omega-3-fatty acids, non-steroidalanti-inflammtories (NSAIDs) and/or opioids.

By “administered in combination” it is meant to include simultaneousadministration in a single formulation, simultaneous administration inseparate formulations, sequential administration of the compositioncomprising egg yolk powder followed by the one or more additional agentsand vice versa, and administration of separate formulations at separatetimes.

The compositions described herein can be formulated for administrationto a subject via any conventional means including, but not limited to,oral, or buccal.

Moreover, the compositions described herein, can be formulated into anysuitable dosage form, including but not limited to, aqueous oraldispersions, liquids, gels, syrups, elixirs, slurries, suspensions andthe like, for oral ingestion by an individual in need, solid oral dosageforms, controlled release formulations, fast melt formulations,effervescent formulations, lyophilized formulations, tablets, powders,pills, dragees, capsules, delayed release formulations, aqueous liquiddispersions, self-emulsifying dispersions, solid solutions, liposomaldispersions, solid dosage forms, powders, tablets, capsules, pills,delayed release formulations.

Formulations for oral use can be obtained by mixing one or more solidexcipient with one or more of the compounds described herein, optionallygrinding the resulting mixture, and processing the mixture of granules,after adding suitable auxiliaries, if desired, to obtain tablets ordragee cores. Suitable excipients include, for example, fillers such assugars, including glucose, fructose, lactose, sucrose, mannitol,sorbitol, stevia extract, or sucralose; cellulose preparations such as,for example, maize starch, wheat starch, rice starch, potato starch,gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose,hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or otherssuch as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. Ifdesired, disintegrating agents may be added, such as the cross-linkedcroscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or asalt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Formulations which can be used orally include push-fit capsules made ofgelatin, as well as soft, sealed capsules made of gelatin and aplasticizer, such as glycerol or sorbitol. The push-fit capsules cancontain the active ingredients in admixture with fillers such aslactose, binders such as starches, and/or lubricants such as talc ormagnesium stearate and, optionally, stabilizers. In soft capsules, theactive compounds may be dissolved or suspended in suitable liquids, suchas fatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added. All formulations for oraladministration should be in dosages suitable for such administration.

In some embodiments, the solid dosage forms disclosed herein may be inthe form of a tablet, (including a suspension tablet, a fast-melttablet, a bite-disintegration tablet, a rapid-disintegration tablet, aneffervescent tablet, or a caplet), a pill, a powder (including a sterilepackaged powder, a dispensable powder, or an effervescent powder) acapsule (including both soft or hard capsules, e.g., capsules made fromanimal-derived gelatin or plant-derived HPMC, or “sprinkle capsules”),solid dispersion, solid solution, pellets, granules. In otherembodiments, the pharmaceutical formulation is in the form of a powder.In still other embodiments, the pharmaceutical formulation is in theform of a tablet. Additionally, formulations described herein may beadministered as a single capsule or in multiple capsule dosage form. Insome embodiments, the formulation is administered in two, or three, orfour, capsules or tablets.

Soft gel or soft gelatin capsules may be prepared, for example, withoutlimitation, by dispersing the formulation in an appropriate vehicle(vegetable oils are commonly used) to form a high viscosity mixture.This mixture is then encapsulated with a gelatin-based film usingtechnology and machinery known to those in the soft gel industry. Theindustrial units so formed are then dried to constant weight.

In some embodiments, the formulations may include other medicinal orpharmaceutical agents, carriers, diluents, dispersing agents, suspendingagents, thickening agents, adjuvants, such as preserving, stabilizing,wetting or emulsifying agents, solution promoters, and/or buffers. Inaddition, the formulations can also contain other therapeuticallyvaluable substances.

The formulations described herein can include egg yolk powder and one ormore pharmaceutically acceptable additives such as a compatible carrier,binder, filling agent, suspending agent, flavoring agent, sweeteningagent, disintegrating agent, dispersing agent, surfactant, lubricant,colorant, diluent, solubilizer, moistening agent, plasticizer,stabilizer, penetration enhancer, wetting agent, anti-foaming agent,antioxidant, preservative, or one or more combination(s) thereof. Instill other aspects, using standard coating procedures, a film coatingis provided around the formulation of the compound described herein. Inone embodiment, some or all of the particles of the compound describedherein are coated. In another embodiment, some or all of the particlesof the compound described herein are microencapsulated. In still anotherembodiment, the particles of the compound described herein are notmicroencapsulated and are uncoated.

In certain embodiments, compositions may also include one or more pHadjusting agents or buffering agents, including acids such as acetic,boric, citric, lactic, phosphoric and hydrochloric acids; bases such assodium hydroxide, sodium phosphate, sodium borate, sodium citrate,sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; andbuffers such as citrate/dextrose, sodium bicarbonate and ammoniumchloride. Such acids, bases and buffers are included in an amountrequired to maintain pH of the composition in an acceptable range.

In other embodiments, compositions may also include one or more salts inan amount required to bring osmolality of the composition into anacceptable range. Such salts include those having sodium, potassium orammonium cations and chloride, citrate, ascorbate, borate, phosphate,bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable saltsinclude sodium chloride, potassium chloride, sodium thiosulfate, sodiumbisulfite and ammonium sulfate.

Formulations including egg yolk powder, as described herein, may bemanufactured in a conventional manner, such as, by way of example only,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

In certain embodiments, compositions provided herein may also includeone or more preservatives to inhibit microbial activity. Suitablepreservatives include mercury-containing substances such as merfen andthiomersal; stabilized chlorine dioxide; and quaternary ammoniumcompounds such as benzalkonium chloride, cetyltrimethylammonium bromideand cetylpyridinium chloride.

Formulations described herein may benefit from antioxidants, metalchelating agents, thiol containing compounds and other generalstabilizing agents. Examples of such stabilizing agents, include, butare not limited to: (a) about 0.5% to about 2% w/v glycerol, (b) about0.1% to about 1% w/v methionine, (c) about 0.1% to about 2% w/vmonothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% toabout 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v polysorbate80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h) arginine, (i)heparin, (j) dextran sulfate, (k) cyclodextrins, (l) pentosanpolysulfate and other heparinoids, (m) divalent cations such asmagnesium and zinc; or (n) combinations thereof.

Binders imparting cohesive qualities may also be used. Examples include,but are not limited to, alginic acid and salts thereof; cellulosederivatives such as carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose,hydroxypropylcellulose, ethylcellulose, and microcrystalline cellulose;microcrystalline dextrose; amylose; magnesium aluminum silicate;polysaccharide acids; bentonites; gelatin; polyvinylpyrrolidone/vinylacetate copolymer; crosspovidone; povidone; starch; pregelatinizedstarch; tragacanth, dextrin, a sugar, such as sucrose, glucose,dextrose, molasses, mannitol, sorbitol, xylitol, and lactose; a naturalor synthetic gum such as acacia, tragacanth, ghatti gum, mucilage ofisapol husks, polyvinylpyrrolidone, larch arabogalactan, polyethyleneglycol, waxes, sodium alginate, and the like.

In general, binder levels of 20-70% are used in powder-filled gelatincapsule formulations. Binder usage level in tablet formulations varieswhether direct compression, wet granulation, roller compaction, or usageof other excipients such as fillers which itself can act as moderatebinder.

Formulators skilled in art can determine the binder level for theformulations, but binder usage level of up to 70% in tablet formulationsis common.

Compositions may further comprise carriers of relatively nontoxicchemical compounds or agents that facilitate the incorporation of acompound into cells or tissues. Nonlimiting examples include binders,suspending agents, disintegration agents, filling agents, surfactants,solubilizers, stabilizers, lubricants, wetting agents, diluents, and thelike. Suitable carriers for use in solid dosage forms described hereininclude, but are not limited to, acacia, gelatin, colloidal silicondioxide, calcium glycerophosphate, calcium lactate, maltodextrin,glycerine, magnesium silicate, sodium caseinate, soy lecithin, sodiumchloride, tricalcium phosphate, dipotassium phosphate, sodium stearoyllactylate, carrageenan, monoglyceride, diglyceride, pregelatinizedstarch, hydroxypropylmethylcellulose, hydroxypropylmethylcelluloseacetate stearate, sucrose, microcrystalline cellulose, lactose, mannitoland the like.

Dispersing agents and/or viscosity modulating agents include materialsthat control the diffusion and homogeneity of a compound through liquidmedia or a granulation method or blend method. In some embodiments,these agents also facilitate the effectiveness of a coating or erodingmatrix. Nonlimiting examples of diffusion facilitators/dispersing agentsinclude hydrophilic polymers, electrolytes, a Tween, PEG,polyvinylpyrrolidone, and carbohydrate-based dispersing agents such ashydroxypropyl celluloses (e.g., HPC, HPC-SL, and HPC-L), hydroxypropylmethylcelluloses (e.g., HPMC K100, HPMC K4M, HPMC K15M, and HPMC K100M),carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose,hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), vinyl pyrrolidone/vinyl acetate copolymer (S630),4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethylene oxide andformaldehyde (also known as tyloxapol), poloxamers, block copolymers ofethylene oxide and propylene oxide; and poloxamines, tetrafunctionalblock copolymers derived from sequential addition of propylene oxide andethylene oxide to ethylenediamine, polyvinylpyrrolidone K12,polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, orpolyvinylpyrrolidone K30, polyvinylpyrrolidone/vinyl acetate copolymer(S-630), polyethylene glycol, e.g., the polyethylene glycol can have amolecular weight of about 300 to about 6000, or about 3350 to about4000, or about 7000 to about 5400, sodium carboxymethylcellulose,methylcellulose, polysorbate-80, sodium alginate, gums, such as, e.g.,gum tragacanth and gum acacia, guar gum, xanthans, including xanthangum, sugars, cellulosics, such as, e.g., sodium carboxymethylcellulose,methylcellulose, sodium carboxymethylcellulose, polysorbate-80, sodiumalginate, polyethoxylated sorbitan monolaurate, polyethoxylated sorbitanmonolaurate, povidone, carbomers, polyvinyl alcohol (PVA), alginates,chitosans and combinations thereof. Plasticizers such as cellulose ortriethyl cellulose can also be used as dispersing agents. Dispersingagents that are particularly useful in liposomal dispersions andself-emulsifying dispersions are dimyristoyl phosphatidyl choline,natural phosphatidyl choline from eggs, natural phosphatidyl glycerolfrom eggs, cholesterol and isopropyl myristate.

Combinations of one or more erosion facilitator with one or morediffusion facilitator can also be used in the present compositions.

Compositions of the present invention may further comprise diluents usedto dilute the compound of interest prior to delivery. Diluents can alsobe used to stabilize compounds because they can provide a more stableenvironment. Salts dissolved in buffered solutions (which also canprovide pH control or maintenance) are utilized as diluents in the art,including, but not limited to a phosphate buffered saline solution. Incertain embodiments, diluents increase bulk of the composition tofacilitate compression or create sufficient bulk for homogenous blendfor capsule filling. Such compounds include e.g., lactose, starch,mannitol, sorbitol, dextrose, microcrystalline cellulose; dibasiccalcium phosphate, dicalcium phosphate dihydrate; tricalcium phosphate,calcium phosphate; anhydrous lactose, spray-dried lactose;pregelatinized starch, compressible sugar; mannitol,hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetatestearate, sucrose-based diluents, confectioner's sugar; monobasiccalcium sulfate monohydrate, calcium sulfate dihydrate; calcium lactatetrihydrate, dextrates; hydrolyzed cereal solids, amylose; powderedcellulose, calcium carbonate; glycine, kaolin; sodium chloride;inositol, bentonite, and the like.

Compositions may further comprise an enteric coating, a substance thatremains substantially intact in the stomach but dissolves and releasesthe egg yolk powder in the small intestine or colon. Generally, theenteric coating comprises a polymeric material that prevents release inthe low pH environment of the stomach but that ionizes at a higher pH,typically a pH of 6 to 7, and thus dissolves sufficiently in the smallintestine or colon to release the active agent therein.

In addition, the compositions may comprise an erosion facilitator, amaterial that controls the erosion of a particular material ingastrointestinal fluid. Erosion facilitators are generally known tothose of ordinary skill in the art. Exemplary erosion facilitatorsinclude, e.g., hydrophilic polymers, electrolytes, proteins, peptides,and amino acids.

Filling agents including compounds such as lactose, calcium carbonate,calcium phosphate, dibasic calcium phosphate, calcium sulfate,microcrystalline cellulose, cellulose powder, dextrose, dextrates,dextran, starches, pregelatinized starch, sucrose, xylitol, lactitol,mannitol, sorbitol, sodium chloride, polyethylene glycol, and the likecan also be included in the compositions. Suitable filling agents foruse in the solid dosage forms described herein include, but are notlimited to, lactose, calcium carbonate, calcium phosphate, dibasiccalcium phosphate, calcium sulfate, microcrystalline cellulose,cellulose powder, dextrose, dextrates, dextran, starches, pregelatinizedstarch, hydroxypropylmethycellulose (HPMC), hydroxypropylmethycellulosephthalate, hydroxypropylmethylcellulose acetate stearate (HPMCAS),sucrose, xylitol, lactitol, mannitol, sorbitol, sodium chloride,polyethylene glycol, and the like.

In addition, flavoring agents and/or sweeteners can be used in thecompositions and may include acacia syrup, acesulfame K, alitame, anise,apple, aspartame, banana, Bavarian cream, berry, black currant,butterscotch, calcium citrate, camphor, caramel, cherry, cherry cream,chocolate, cinnamon, bubble gum, citrus, citrus punch, citrus cream,cotton candy, cocoa, cola, cool cherry, cool citrus, cyclamate,cylamate, dextrose, eucalyptus, eugenol, fructose, fruit punch, ginger,glycyrrhetinate, glycyrrhiza (licorice) syrup, grape, grapefruit, honey,isomalt, lemon, lime, lemon cream, monoammonium glyrrhizinate, maltol,mannitol, maple, marshmallow, menthol, mint cream, mixed berry,neohesperidine DC, neotame, orange, pear, peach, peppermint, peppermintcream, raspberry, root beer, rum, saccharin, safrole, sorbitol,spearmint, spearmint cream, strawberry, strawberry cream, stevia,sucralose, sucrose, sodium saccharin, saccharin, aspartame, acesulfamepotassium, mannitol, talin, sylitol, sucralose, sorbitol, Swiss cream,tagatose, tangerine, thaumatin, tutti fruitti, vanilla, walnut,watermelon, wild cherry, wintergreen, xylitol, or any combination ofthese flavoring ingredients, e.g., anise-menthol, cherry- anise,cinnamon-orange, cherry-cinnamon, chocolate-mint, honey-lemon,lemon-lime, lemon-mint, menthol-eucalyptus, orange-cream, vanilla-mint,and mixtures thereof.

The compositions may further comprise lubricants and/or glidants thatprevent, reduce or inhibit adhesion or friction of materials.Nonlimiting examples of lubricants include stearic acid, calciumhydroxide, talc, sodium stearyl fumerate, a hydrocarbon such as mineraloil, or hydrogenated vegetable oil such as hydrogenated soybean oil,higher fatty acids and their alkali-metal and alkaline earth metalsalts, such as aluminum, calcium, magnesium, zinc, stearic acid, sodiumstearates, glycerol, talc, waxes, boric acid, sodium benzoate, sodiumacetate, sodium chloride, leucine, a polyethylene glycol (e.g.,PEG-4000) or a methoxypolyethylene glycol, sodium oleate, sodiumbenzoate, glyceryl behenate, polyethylene glycol, magnesium or sodiumlauryl sulfate, colloidal silica, a starch such as corn starch, siliconeoil, a surfactant, and the like.

Plasticizers, compounds used to soften the microencapsulation materialor film coatings to make them less brittle may also be included in thecompositions. Examples of suitable plasticizers include, but are notlimited to, polyethylene glycols such as PEG 300, PEG 400, PEG 600, PEG1450, PEG 3350, and PEG 800, stearic acid, propylene glycol, oleic acid,triethyl cellulose and triacetin. In some embodiments, plasticizers canalso function as dispersing agents or wetting agents.

The compositions may further comprise solubilizers such as triacetin,triethylcitrate, ethyl oleate, ethyl caprylate, sodium lauryl sulfate,sodium doccusate, vitamin E TPGS, dimethylacetamide,N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone,hydroxypropylmethyl cellulose, hydroxypropyl cyclodextrins, ethanol,n-butanol, isopropyl alcohol, cholesterol, bile salts, polyethyleneglycol 200-600, glycofurol, transcutol, propylene glycol, and dimethylisosorbide and the like.

In addition, the compositions my comprise stabilizers such asantioxidation agents, buffers, acids, preservatives and the like.

Suitable suspending agents for use in solid dosage forms described hereinclude, but are not limited to, polyvinylpyrrolidone, e.g.,polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyethylene glycol, e.g., thepolyethylene glycol can have a molecular weight of about 300 to about6000, or about 3350 to about 4000, or about 7000 to about 5400, vinylpyrrolidone/vinyl acetate copolymer (S630), sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as,e.g., gum tragacanth and gum acacia, guar gum, xanthans, includingxanthan gum, sugars, cellulosics, such as, e.g., sodiumcarboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80,sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylatedsorbitan monolaurate, povidone and the like.

Surfactants including compounds such as sodium lauryl sulfate, sodiumdocusate, Tweens, triacetin, vitamin E TPGS, sorbitan monooleate,polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bilesalts, glyceryl monostearate, copolymers of ethylene oxide and propyleneoxide and the like may also be included. Additional surfactants includepolyoxyethylene fatty acid glycerides and vegetable oils, e.g.,polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylenealkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40. Insome embodiments, surfactants may be included to enhance physicalstability or for other purposes.

Viscosity enhancing agents including, e.g., methyl cellulose, xanthangum, carboxymethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetatestearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinylalcohol, alginates, acacia, chitosans and combinations thereof may alsobe included.

In addition, wetting agents including compounds such as oleic acid,glyceryl monostearate, sorbitan monooleate, sorbitan monolaurate,triethanolamine oleate, polyoxyethylene sorbitan monooleate,polyoxyethylene sorbitan monolaurate, sodium docusate, sodium oleate,sodium lauryl sulfate, sodium doccusate, triacetin, Tween 80, vitamin ETPGS, ammonium salts and the like may be included in these compositions.

In some embodiments, solid dosage forms, e.g., tablets, capsules, areprepared by mixing the egg yolk powder described herein, with one ormore pharmaceutical excipients to form a bulk blend composition. Whenreferring to these bulk blend compositions as homogeneous, it is meantthat the particles of egg yolk powder, are dispersed evenly throughoutthe composition so that the composition may be readily subdivided intoequally effective unit dosage forms, such as tablets, pills, andcapsules.

Conventional techniques include, e.g., one or a combination of methods:(1) dry mixing, (2) direct compression, (3) milling, (4) dry ornon-aqueous granulation, (5) wet granulation, or (6) fusion. See, e.g.,Lachman et al., “The Theory and Practice of Industrial Pharmacy” (1986).

It should be appreciated that there is considerable overlap betweenadditives used in the solid dosage forms described herein. Thus, theabove-listed additives should be taken as merely exemplary, and notlimiting, of the types of additives that can be included.

A capsule may be prepared, for example, by placing the bulk blend of theformulation of the compound described above, inside of a capsule. Insome embodiments, the formulations (non-aqueous suspensions andsolutions) are placed in a soft gelatin capsule. In other embodiments,the formulations are placed in standard gelatin capsules or non-gelatincapsules such as capsules comprising HPMC. In other embodiments, theformulation is placed in a sprinkle capsule, wherein the capsule may beswallowed whole or the capsule may be opened and the contents sprinkledon food prior to eating. In some embodiments, the therapeutic dose issplit into multiple (e.g., two, three, or four) capsules. In someembodiments, the entire dose of the formulation is delivered in acapsule form.

In another aspect, dosage forms may include microencapsulatedformulations. In some embodiments, one or more other compatiblematerials are present in the microencapsulation material. Exemplarymaterials include, but are not limited to, pH modifiers, erosionfacilitators, anti-foaming agents, antioxidants, flavoring agents, andcarrier materials such as binders, suspending agents, disintegrationagents, filling agents, surfactants, solubilizers, stabilizers,lubricants, wetting agents, and diluents.

Materials useful for the microencapsulation described herein includematerials which sufficiently isolate the compound from othernon-compatible excipients. Materials compatible with the egg yolk powderare those that delay the release of the egg yolk powder in vivo.

In other embodiments, the formulations described herein, which includethe egg yolk powder, are solid dispersions. Methods of producing suchsolid dispersions are known in the art and include, but are not limitedto, for example, U.S. Pat. Nos. 4,343,789, 5,340,591, 5,456,923,5,700,485, 5,723,269, and U.S. Pub. Appl 2004/0013734.

In still other embodiments, the formulations described herein are solidsolutions. Solid solutions incorporate a substance together with theactive agent and other excipients such that heating the mixture resultsin dissolution of the drug and the resulting composition is then cooledto provide a solid blend which can be further formulated or directlyadded to a capsule or compressed into a tablet. Methods of producingsuch solid solutions are known in the art and include, but are notlimited to, for example, U.S. Pat. Nos. 4,151,273, 5,281,420, and6,083,518.

In some embodiments, the solid dosage forms described herein can beformulated as enteric coated delayed release oral dosage forms, i.e., asan oral dosage form of a pharmaceutical composition as described hereinwhich utilizes an enteric coating to affect release in the smallintestine of the gastrointestinal tract. The enteric coated dosage formmay be a compressed or molded or extruded tablet/mold (coated oruncoated) containing granules, powder, pellets, beads or particles ofthe active ingredient and/or other composition components. The entericcoated oral dosage form may also be a capsule (coated or uncoated)containing pellets, beads or granules of the solid carrier or thecomposition.

The term “delayed release” as used herein refers to the delivery so thatthe release can be accomplished at some generally predictable locationin the intestinal tract more distal to that which would have beenaccomplished if there had been no delayed release alterations. In someembodiments the method for delay of release is coating. Any coatingsshould be applied to a sufficient thickness such that the entire coatingdoes not dissolve in the gastrointestinal fluids at pH below about 5,but does dissolve at pH about 5 and above. It is expected that anyanionic polymer exhibiting a pH-dependent solubility profile can be usedas an enteric coating for the methods and compositions described hereinto achieve delivery to the lower gastrointestinal tract.

In some embodiments, formulations are provided that include particles ofegg yolk powder described herein and at least one dispersing agent orsuspending agent for oral administration to a subject. The formulationsmay be a powder and/or granules for suspension, and upon admixture withwater, a substantially uniform suspension is obtained.

Liquid formulation dosage forms for oral administration can be aqueoussuspensions selected from the group including, but not limited to,pharmaceutically acceptable aqueous oral dispersions, emulsions,solutions, elixirs, gels, and syrups. See, e.g., Singh et al.,Encyclopedia of Pharmaceutical Technology, 2nd Ed., pp. 754-757 (2002).In addition to the particles of egg yolk powder, the liquid dosage formsmay include additives, such as: (a) disintegrating agents; (b)dispersing agents; (c) wetting agents; (d) at least one preservative,(e) viscosity enhancing agents, (t) at least one sweetening agent, and(g) at least one flavoring agent. In some embodiments, the aqueousdispersions can further include a crystalline inhibitor.

The aqueous suspensions and dispersions described herein can remain in ahomogenous state, as defined in The USP Pharmacists' Pharmacopeia (2005edition, chapter 905), for at least 4 hours. The homogeneity should bedetermined by a sampling method consistent with regards to determininghomogeneity of the entire composition. In one embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 1 minute. In another embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 45 seconds. In yet another embodiment, anaqueous suspension can be re-suspended into a homogenous suspension byphysical agitation lasting less than 30 seconds. In still anotherembodiment, no agitation is necessary to maintain a homogeneous aqueousdispersion.

Suitable preservatives for the aqueous suspensions or dispersionsdescribed herein include, for example, potassium sorbate, parabens(e.g., methylparaben and propylparaben), benzoic acid and its salts,other esters of parahydroxybenzoic acid such as butylparaben, alcoholssuch as ethyl alcohol or benzyl alcohol, phenolic compounds such asphenol, or quaternary compounds such as benzalkonium chloride.Preservatives, as used herein, are incorporated into the dosage form ata concentration sufficient to inhibit microbial growth.

In one nonlimiting embodiment, the aqueous liquid dispersion cancomprise a sweetening agent or flavoring agent in a concentrationranging from about 0.005% to about 0.5% the volume of the aqueousdispersion. In yet another embodiment, the aqueous liquid dispersion cancomprise a sweetening agent or flavoring agent in a concentrationranging from about 0.01% to about 1.0% the volume of the aqueousdispersion.

In addition to the additives listed above, the liquid formulations canalso include inert diluents commonly used in the art, such as water orother solvents, solubilizing agents, and emulsifiers. Exemplaryemulsifiers are ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propyleneglycol,1,3-butyleneglycol, dimethylformamide, sodium lauryl sulfate, sodiumdoccusate, cholesterol, cholesterol esters, taurocholic acid,phosphotidylcholine, oils, such as cottonseed oil, groundnut oil, corngerm oil, olive oil, castor oil, and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters ofsorbitan, or mixtures of these substances, and the like.

In some embodiments, the formulations described herein can beself-emulsifying drug delivery systems (SEDDS). Emulsions aredispersions of one immiscible phase in another, usually in the form ofdroplets. Generally, emulsions are created by vigorous mechanicaldispersion.

SEDDS, as opposed to emulsions or microemulsions, spontaneously formemulsions when added to an excess of water without any externalmechanical dispersion or agitation. An advantage of SEDDS is that onlygentle mixing is required to distribute the droplets throughout thesolution. Additionally, water or the aqueous phase can be added justprior to administration, which ensures stability of an unstable orhydrophobic active ingredient. Thus, the SEDDS provides an effectivedelivery system for oral and parenteral delivery of hydrophobic activeingredients. SEDDS may provide improvements in the bioavailability ofhydrophobic active ingredients. Methods of producing self-emulsifyingdosage forms are known in the art and include, but are not limited to,for example, U.S. Pat. Nos. 5,858,401, 6,667,048, and 6,960,563.

Buccal formulations that include egg yolk powder may be administeredusing a variety of formulations known in the art. For example, suchformulations include, but are not limited to, U.S. Pat. Nos. 4,229,447,4,596,795, 4,755,386, and 5,739,136. In addition, the buccal dosageforms described herein can further include a bioerodible (hydrolysable)polymeric carrier that also serves to adhere the dosage form to thebuccal mucosa. The buccal dosage form is fabricated so as to erodegradually over a predetermined time period. Buccal drug delivery, aswill be appreciated by those skilled in the art, avoids thedisadvantages encountered with oral drug administration, e.g., slowabsorption, degradation of the active agent by fluids present in thegastrointestinal tract and/or first-pass inactivation in the liver. Withregard to the bioerodible (hydrolysable) polymeric carrier, it will beappreciated that virtually any such carrier can be used, so long as thedesired drug release profile is not compromised, and the carrier iscompatible with the egg yolk powder, and any other components that maybe present in the buccal dosage unit. Generally, the polymeric carriercomprises hydrophilic (water-soluble and water-swellable) polymers thatadhere to the wet surface of the buccal mucosa. Other components mayalso be incorporated into the buccal dosage forms described hereininclude, but are not limited to, disintegrants, diluents, binders,lubricants, flavoring, colorants, preservatives, and the like. Forbuccal or sublingual administration, the compositions may take the formof tablets, lozenges, or gels formulated in a conventional manner.

In certain embodiments, delivery systems for pharmaceutical compoundsmay be employed, such as, for example, liposomes and emulsions. Incertain embodiments, compositions provided herein can also include amucoadhesive polymer, selected from among, for example,carboxymethylcellulose, carbomer (acrylic acid polymer),poly(methylmethacrylate), polyacrylamide, polycarbophil, acrylicacid/butyl acrylate copolymer, sodium alginate and dextran.

The invention is further illustrated by the following example, whichshould not be construed as further limiting. The contents of allreferences, pending patent applications, and published patents citedthroughout this application are hereby expressly incorporated byreference.

EXAMPLE

100 dogs with cranial cruciate ligament rupture were enrolled in thisstudy. Dogs were randomized to receive either FORTETROPIN or a placeboin a double-blind fashion. Dogs were given 300 mg/kg/day of eitherFORTETROPIN or placebo starting a week prior to TPLO surgery andcontinuing for 12 weeks post-surgery. At pre-surgery and 6 and 12 weekspost-surgery, thigh circumference, thickness of coxofemoral extensorsand flexors, thickness of epaxial muscle, were measured. In addition,serum myostatin and C-reactive protein levels were measured and stanceanalysis was performed on each dog at pre-surgery and again 12-weekspost-surgery.

Dogs that received FORTETROPIN experienced a smaller decrease in thighcircumference over 8 weeks relative to dogs that received a placebo.

Dogs that received FORTETROPIN also experienced a smaller increase inmyostatin concentration over 8 weeks relative to dogs that received aplacebo.

In addition, dogs that received FORTETROPIN experienced a greaterincrease in force that could be exerted due to weight on the affectedhind limb after 8 weeks relative to placebo.

What is claimed is:
 1. A method for inhibiting, reversing and/oralleviating muscle disuse atrophy in a mammal, said method comprisingadministering to the mammal a composition comprising egg yolk powder. 2.The method of claim 1 wherein the composition comprises a fertilized eggyolk derived product.
 3. The method of claim 2 wherein the fertilizedegg yolk derived product is FORTETROPIN.
 4. The method of claim 1wherein the egg yolk powder is administered in an amount effective toupregulate mTor pathway activity, downregulate ubiquitin proteasomepathway activity, downregulate serum myostatin levels and/or reduceActRIIB expression.
 5. The method of claim 1 wherein the compositioncomprises avian follistatin.
 6. The method of claim 1 wherein thecomposition is administered to the mammal prior to a surgical procedure,following a surgical procedure or prior to and following a surgicalprocedure.
 7. The method of claim 6 wherein the surgical procedure is anorthopedic surgical procedure.
 8. The method of claim 1 wherein thecomposition is administered to the mammal following an immobilizinginjury.
 9. The method of claim 1 wherein administration of thecompositions results in reduced muscle atrophy of disuse, increasedmuscle thickness, lower reduction in muscle thickness and/or musclemass, improved muscle function, increased muscle force generation,reduced inflammation, reduced recovery time and/or lower increases inmyostatin in the mammal.
 10. The method of claim 1 further comprisingadministering one or more additional agents which alleviate pain and/orinflammation post surgery.